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酶联免疫吸附测定法检测灭活疫苗接种及后续攻毒后猪血清中抗猪繁殖与呼吸综合征病毒抗体的能力。

Ability of ELISAs to detect antibodies against porcine respiratory and reproductive syndrome virus in serum of pigs after inactivated vaccination and subsequent challenge.

作者信息

Sattler Tatjana, Pikalo Jutta, Wodak Eveline, Schmoll Friedrich

机构信息

Large Animal Clinic for Internal Medicine, University of Leipzig, An den Tierkliniken 11, 04103, Leipzig, Germany.

Institute for Veterinary Disease Control, AGES, Robert-Koch-Gasse 17, 2340, Mödling, Austria.

出版信息

BMC Vet Res. 2016 Nov 21;12(1):259. doi: 10.1186/s12917-016-0888-0.

Abstract

BACKGROUND

In this study, six enzyme-linked immunosorbent assays (ELISA), intended for routine porcine reproductive and respiratory syndrome virus (PRRSV) herd monitoring, are tested for their ability to detect PRRSV specific antibodies in the serum of pigs after vaccination with an inactivated PRRSV type 1 vaccine and subsequent infection with a highly pathogenic (HP) PRRSV field strain. For this reason, ten piglets (group V) from a PRRSV negative herd were vaccinated twice at the age of 2 and 4 weeks with an inactivated PRRSV vaccine. Ten additional piglets (group N) from the same herd remained unvaccinated. Three weeks after second vaccination, each of the piglets received an intradermal application of an HP PRRSV field strain. Serum samples were taken before first vaccination as well as before and 3, 7, 10 and 14 days after HP PRRSV application. All serum samples were tested for PRRSV RNA by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) as well as for PRRSV antibodies with all six study ELISAs.

RESULTS

At the beginning of the study (before vaccination), all of the piglets were PRRSV antibody negative with all study ELISAs. They also tested negative for PRRSV RNA measured by RT-qPCR. From day 3 after HP PRRSV application until the end of the study, a viremia was detected by RT-qPCR in all of the piglets. On day 0 (day of HP PRRSV application), nine out of ten piglets of the pre-vaccinated group tested PRRSV antibody positive with one of the tested ELISAs, although with lower S/P values than after infection. On day 10 after HP PRRSV application, all study ELISAs except one had significantly higher S/P or OD values, respectively more positive samples, in group V than in group N.

CONCLUSIONS

Only one of the tested ELISAs was able to detect reliably PRRSV antibodies in pigs vaccinated with an inactivated PRRSV vaccine. With most of the tested ELISAs, higher S/P values respectively more positive samples after PRRSV infection were seen in the pre-vaccinated group than in the non-vaccinated.

摘要

背景

在本研究中,对六种用于猪繁殖与呼吸综合征病毒(PRRSV)常规群体监测的酶联免疫吸附测定(ELISA)进行了测试,以评估其检测接种1型灭活PRRSV疫苗后再感染高致病性(HP)PRRSV田间毒株的猪血清中PRRSV特异性抗体的能力。为此,从一个PRRSV阴性猪群中选取十头仔猪(V组),在2周龄和4周龄时用1型灭活PRRSV疫苗进行两次接种。同一猪群中另外十头仔猪(N组)未接种疫苗。第二次接种三周后,每头仔猪皮内接种一株HP PRRSV田间毒株。在首次接种前以及接种HP PRRSV之前、之后3、7、10和14天采集血清样本。所有血清样本均通过逆转录定量聚合酶链反应(RT-qPCR)检测PRRSV RNA,并使用所有六种研究用ELISA检测PRRSV抗体。

结果

在研究开始时(接种疫苗前),所有仔猪的所有研究用ELISA检测PRRSV抗体均为阴性。RT-qPCR检测PRRSV RNA也呈阴性。从接种HP PRRSV后第3天直至研究结束,所有仔猪经RT-qPCR检测均出现病毒血症。在接种HP PRRSV的当天(第0天),接种疫苗前的十头仔猪中有九头用其中一种检测ELISA检测PRRSV抗体呈阳性,尽管S/P值低于感染后。接种HP PRRSV后第10天,除一种ELISA外,所有研究用ELISA在V组中的S/P值或OD值均显著更高,阳性样本更多。

结论

所测试的ELISA中只有一种能够可靠地检测接种1型灭活PRRSV疫苗的猪中的PRRSV抗体。对于大多数测试的ELISA,接种疫苗前的组在PRRSV感染后的S/P值更高,阳性样本更多。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/22c6/5117522/d59189ac7424/12917_2016_888_Fig1_HTML.jpg

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